Abstract
Glucose-responsive formulations of insulin can increase its therapeutic index and reduce the burden of its administration. However, it has been difficult to develop single-dosage formulations that can release insulin in both a sustained and glucose-responsive manner. Here we report the development of a subcutaneously injected glucose-responsive formulation that nearly does not trigger the formation of a fibrous capsule and that leads to week-long normoglycaemia and negligible hypoglycaemia in mice and minipigs with type 1 diabetes. The formulation consists of gluconic acid-modified recombinant human insulin binding tightly to poly-l-lysine modified by 4-carboxy-3-fluorophenylboronic acid via glucose-responsive phenylboronic acid–diol complexation and electrostatic attraction. When the insulin complex is exposed to high glucose concentrations, the phenylboronic acid moieties of the polymers bind rapidly to glucose, breaking the complexation and reducing the polymers’ positive charge density, which promotes the release of insulin. The therapeutic performance of this long-acting single-dose formulation supports its further evaluation and clinical translational studies.
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Data availability
The authors declare that all of the data supporting the findings of this study are available within the paper and its Supplementary Information. Source data are provided with this paper.
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Acknowledgements
This work was supported by the grants from National Key R&D Program of China (grant no. 2022YFE0202200), Juvenile Diabetes Research Foundation (grant nos 2-SRA-2021-1064-M-B, 2-SRA-2022-1159-M-B), and Zhejiang University’s start-up packages, the Kunpeng Program grant and Fundamental Research Funds for the Central Universities (grant no. 2021FZZX001-46). A.R.K. is supported by the National Center for Advancing Translational Sciences, National Institutes of Health, through grant no. KL2TR002490. The project was supported by grant no. UL1TR002489 from the Clinical and Translational Science Award programme of the National Center for Advancing Translational Sciences, National Institutes of Health. We appreciate the help from G. Zhu and Y. Zhang (Cryo-EM Centre, Zhejiang University) for processing of samples for electron microscopy, D. Guo (Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine) for slicing and staining skin tissue, C. Sun (Analysis Center of Agrobiology and Environmental Sciences, Zhejiang University) for MALDI-TOF mass analysis and D. Xu (Animal Center, Zhejiang University) for taking care of the minipigs.
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Z.G., J.B.B. and J.W. conceived and designed the study. J.W., J.Z., X.W., W.L., Y.W., W.Z., T.S., Y.Z., Y.L., K.J. and J.X. conducted experiments and obtained data. X.Z. and P.Z. provided experimental and theoretical guidance, respectively. H.Z. and Y.X. conducted western blot experiments and provided theoretical support, respectively. J.W., J.Z., X.W., W.L., Y.Z., W.Z., T.S., J.B.B., A.R.K. and Z.G. analysed the data and wrote the paper.
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Z.G. is the co-founder of Zenomics Inc., Zcapsule Inc. and μZen Inc. The remaining authors declare no competing interests.
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Extended data
Extended Data Fig. 1 BG regulation by the insulin complex in diabetic mice.
The dose was set to 2.5 (a), 5 (b), 10 (c), and 20 (d) mg/kg. Insulin complex was subcutaneously injected. Data points are means ± SD (n = 5).
Extended Data Fig. 2 Time proportion in the range of hyperglycaemia, normoglycaemia and hypoglycaemia in diabetic minipigs after subcutaneous injection of the complex or of insulin glargine.
Hyperglycaemia, normoglycaemia, and hypoglycaemia were defined as BG more than 200 mg/dL, 50-200 mg/dL, and less than 50 mg/dL. Data points are means ± SD (n = 3). a, Both insulin glargine (0.4 to 0.6 U/kg) and complex (0.2 to 0.3 mg/kg) were injected once at the beginning of treatment. As BG was monitored for 3 days for the insulin-treated group, BG within three days was included in the statistical analysis. b, Insulin glargine (0.4 to 0.6 U/kg) was injected daily for 7 days, and complex (0.2 to 0.3 mg/kg) was injected once at the beginning of treatment. BG within 7 days was included in the statistical analysis. Two-tailed Student’s t-test was used for statistical analysis.
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Supplementary Figs. 1–13.
Supplementary Data
Source data for Supplementary Figs. 2 and 3 and 6–9.
Supplementary Data
Unprocessed western blots for Supplementary Fig. 2d.
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Source Data
Statistical source data for Figs. 1–5 and Extended Data Figs. 1–2.
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Zhang, J., Wei, X., Liu, W. et al. Week-long normoglycaemia in diabetic mice and minipigs via a subcutaneous dose of a glucose-responsive insulin complex. Nat. Biomed. Eng (2023). https://doi.org/10.1038/s41551-023-01138-7
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DOI: https://doi.org/10.1038/s41551-023-01138-7
